title: Arachidic acid in extender improves post-thaw parameters of cryopreserved Nili-Ravi buffalo bull semen.
abstract: Cryopreservation process reduces lipids and phospholipids from buffalo bull spermatozoa. It was therefore hypothesized that supplementation of fatty acid to extender may improve the post-thaw quality of buffalo semen. The objective was to evaluate the effect of arachidic acid supplementation in extender on post-thaw quality of buffalo bull (Bubalus bubalis) spermatozoa. Semen was collected from three adult Nili-Ravi buffalo bulls of similar age group with artificial vagina (42°C) for 3 weeks (replicate). Qualified semen ejaculates (n = 18) were split into four aliquots and diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0 ng/ml at 37°C having approximately 50 × 10(6) spermatozoa/ml. Diluted semen was cooled to 4°C in 2 h and equilibrated for 4 h at 4°C. Cooled semen was filled in 0.5-ml straws at 4°C, kept on liquid nitrogen vapours for 10 min and plunged in liquid nitrogen for storage. Thawing of frozen semen was performed after 24 h at 37°C for 30 s. Sperm progressive motility (%) was improved in a dose-dependent manner by supplementing arachidic acid at 5.0, 10.0 and 20.0 ng/ml compared with control. Structural and functional integrity of sperm plasma membrane (%), number of acrosome-intact live sperm (%) and sperm chromatin integrity (%) were better (p < 0.05) in extender having 5.0 ng/ml of arachidic acid compared with control. At 10.0 ng/ml, these values did not vary (p > 0.05) from those at 5.0 ng/ml. Further improvement in structural and functional integrity of sperm plasma membrane, number of acrosome-intact live sperm and chromatin integrity was observed at 20.0 ng/ml of arachidic acid in extender. In conclusion, arachidic acid supplementation in extender improved the post-thaw quality parameters of cryopreserved Nili-Ravi buffalo bull spermatozoa. Among the arachidic acid concentrations studied, maximum improvement in post-thaw semen quality parameters was observed at 20.0 ng/ml.
authors.affiliation: Department of Zoology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan.
authors.forename: R
authors.initials: R
authors.lastname: Ejaz
authors.forename: M S
authors.initials: MS
authors.lastname: Ansari
authors.forename: B A
authors.initials: BA
authors.lastname: Rakha
authors.forename: N
authors.initials: N
authors.lastname: Ullah
authors.forename: A U
authors.initials: AU
authors.lastname: Husna
authors.forename: R
authors.initials: R
authors.lastname: Iqbal
authors.forename: S
authors.initials: S
authors.lastname: Akhter
mesh_terms: D000177:Acrosome / Q000648:ultrastructure; D000818:Animals; D002020:Buffaloes; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents; D004305:Dose-Response Relationship, Drug; D004537:Eicosanoic Acids / Q000008:administration & dosage; D006358:Hot Temperature; D008297:Male; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure
chemical_list: D002843:Chromatin; D003451:Cryoprotective Agents; D004537:Eicosanoic Acids; C094477:arachidic acid